RE: [Histonet] Hardening very soft brains

From:

"Monson, Frederick "

Morning Steve,
Well, ---____..... Oh, I can't do this. The temptation to
humor is really strong on a Friday, but Ill try to detrain myself from
this sticky morsel.
Thus,
My approach would be to determine the reason for the absence of
hardening.
Two possibilities come to mind.
1. Very high phospholipid content which might interfere
with diffusion of HCHO.
2. Very high GAG content which might leave residual
'stickiness'. I like this better!*
Solutions
Lower the temp at which you handle tissues.
Add GAG 'precipitators' to fixative, i.e.
cetylpyridinium chloride(?)
Use wax paper instead of wrapping paper, it's just more
paraffin.
*I'm overcome with the notion that the cranium is first filled with
ground substance that serves as both nutrient and matrix for later
development. This latter idea may imply to some that I have missed some
parts of the latter stages of development and am, thus, using residual
ground substance for dreaming up such 'curious' suggestions. Of course,
if one spends a lifetime promulgating curious ideas, one is bound to
hear complaints that they are grounded in no substance at all.
Finally, I hope for your sake that John Kiernan is awake and
thinking, with his highly developed substance, about your problem. He is
one who has been trained in that chapter of the Histology texts that is
often skipped for lack of time in the normal courses of histo-events.
Cheers,
Fred Monson
Frederick C. Monson, PhD
Center for Advanced Scientific Imaging
Mail to Geology
West Chester University of Pennsylvania
Schmucker II Science Center, Room SS024
South Church Street and Rosedale Avenue
West Chester, PA, 19383
Phone/FAX: 610-738-0437
eMail: fmonson@wcupa.edu
CASI Page and Scheduling
http://darwin.wcupa.edu/CASI/
-----Original Message-----
From: Steve Machin UK [mailto:stevemachinuk@yahoo.co.uk]
Sent: Friday, December 05, 2003 9:11 AM
To: Histonet Histonet Histonet
Subject: [Histonet] Hardening very soft brains
Could anyone help us with a problem we have in processing very soft
fetal brains?
We currently fix in 20% formalin in buffer but the brains are so soft
after processing they stick to the wrapping paper.
Any ideas on how we can harden them after fixation?
Best Wishes
Steve Machinu UK
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